Exploring Thermophilic Enzymes: From Structural Foundations to Functional Enhancement

A special issue of Biomolecules (ISSN 2218-273X). This special issue belongs to the section "Enzymology".

Deadline for manuscript submissions: 28 February 2026 | Viewed by 295

Special Issue Editor


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Guest Editor
Key Laboratory for Molecular Enzymology and Engineering, The Ministry of Education, School of Life Sciences, Jilin University, Changchun 130012, China
Interests: screening and development of new thermophilic enzymes; molecular modification and immobilization of thermophilic enzymes based on protein engineering; design of thermophilic enzyme reactors, activity regulation, engineering-scale preparation, and catalytic applications

Special Issue Information

Dear Colleagues,

Owing to their excellent stability and functionality at high temperatures, thermophilic enzymes play a crucial role in fields such as biological detection, genetic engineering, food processing, and drug synthesis. This Special Issue aims to collect high-quality original research, reviews, and short communications, with a focus on structural analysis of thermostable enzymes, mechanisms of thermal stability, engineering strategies for functional enhancement, and innovative applications in various environments. By highlighting cutting-edge discoveries and insights, this collection seeks to provide a comprehensive overview of recent advancements, deepening our understanding of the stability-related mechanisms of thermophilic enzymes.

Prof. Dr. Renjun Gao
Guest Editor

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Keywords

  • thermostable enzymes
  • structural biology
  • protein engineering
  • functional enhancement
  • industrial applications
  • thermal stability mechanisms
  • biotechnology

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Published Papers (1 paper)

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Research

16 pages, 3511 KB  
Article
Enhancement of Activity of Thermophilic Inorganic Pyrophosphatase Ton1914 via Site-Directed Mutagenesis
by Siyao Liu, Xinrui Yang, Renjun Gao and Guiqiu Xie
Biomolecules 2025, 15(10), 1395; https://doi.org/10.3390/biom15101395 - 30 Sep 2025
Viewed by 205
Abstract
Inorganic pyrophosphatase (PPase) is an enzyme that catalyzes the hydrolysis of pyrophosphate (PPi) into two phosphates. Ton1914, a thermophilic inorganic pyrophosphatase derived from Thermococcus onnurineus NA1, has good thermal stability and an extremely high optimum temperature and has been shown to reduce pyrophosphate [...] Read more.
Inorganic pyrophosphatase (PPase) is an enzyme that catalyzes the hydrolysis of pyrophosphate (PPi) into two phosphates. Ton1914, a thermophilic inorganic pyrophosphatase derived from Thermococcus onnurineus NA1, has good thermal stability and an extremely high optimum temperature and has been shown to reduce pyrophosphate inhibition. In this study, eight sites were selected based on sequence alignment and software calculations, and multiple single mutants were successfully constructed. After saturation and superposition mutations, six superior mutants were obtained. The enzyme activities of E97Y, D101K and L42F were increased 2.57-, 2.47- and 2.15-fold, respectively, while those of L42F/E97Y, L42F/D101K and E97Y/D101K were increased 2.60-, 2.63- and 1.88-fold, respectively, relative to the wild-type enzyme. Compared to Ton1914, all mutants more effectively increased PCR product quantity, reduced the number of qPCR cycles required to reach the threshold, and improved the efficiency of gene amplification. In the UDP-Galactose (UDP-Gal) synthesis reaction, the addition of mutants could further improve yield. When Ton1914 and mutants with the same activity were added, the yield of UDP-Gal was almost identical, effectively reducing the dosage of pyrophosphatase. Overall, the mutants showed greater prospects for industrial application. Full article
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